Lightning Talk & Poster Australasian Extracellular Vesicles Conference 2018

Generation and comprehensive characterization of exosome-mimetic nanovesicles as an alternative delivery system to exosomes (#58)

Amirmohammad Nasiri Kenari 1 , Kenneth Kastaniegaard 2 , Mitch Shambrook 1 , David Greening 1 , Allan Stensballe 2 , Lesley Cheng 1 , Andrew F Hill 1
  1. Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University, Bundoora, VIC, Australia
  2. Department of Health Science and Technology, Aalborg University, Aalborg, Denmark

The potential of utilizing exosomes as a therapeutic delivery system for biological and chemical drugs is currently a highly active area of clinical research. However, the field is currently facing challenges such as low yield and heterogenicity of exosomes produced by mammalian cells. To address these issues, Mimetic-Nano Vesicles (M-NVs) are an emerging and potentially promising alternative therapeutic delivery system to exosomes. M-NVs can be generated from all types of cell lines using a systematic purification protocol with advantages such as, reproducibility, large scale production, uniformity and cost effectiveness. In this study, M-NVs were generated by extruding Human neuroblastoma SH-SY5Y cells (n = 5) using an Avanti mini-extruder and purified by OptiPrep™ density gradients. Generated M-NVs were comprehensively characterized based on biophysical characteristic and further compared to exosomes.  Our protocol was able to reproducibility demonstrate the production of M-NVs at high-yield (>100-fold) when compared to endogenously secreted exosomes from SH-SY5Y cells. Proteomic analysis indicated the presence of key exosomal membrane surface features on M-NVs. RNA deep sequencing revealed that M-NVs cargo closely mirrored parental cells however, endogenous exosomes were found to specifically package certain RNA species. In summary, results from this study demonstrated a reproducible large-scale production of M-NVs, provided key insights into M-NVs cargo and highlighted its potential as an alternative to exosomes for therapeutic delivery system.